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Novel auto-selection systems for transformation selection of Saccharomyces cerevisiae in rich complex media

Höler, Sebastian ; Bertl, Adam ; Degreif, Daniel (2021):
Novel auto-selection systems for transformation selection of Saccharomyces cerevisiae in rich complex media.
In: FEMS yeast research, 21 (5), ISSN 1567-1364,
DOI: 10.1093/femsyr/foab039,
[Article]

Abstract

The most widely used strategy for selection of yeast transformed with episomal plasmids comprises the use of auxotrophic yeast strains in combination with vectors containing complementing prototrophic marker genes. Another approach uses heterologous genes or cassettes which, if present in the vector, render the otherwise sensitive yeast strain resistant to antibiotics. In addition, auto-selection systems for Saccharomyces cerevisiae have been developed that eliminate the requirement for synthetic drop-out media or the use of antibiotics for transformation selection and subsequent plasmid maintenance in expression cultures. Here we describe a combination of host strain and vector system introducing a novel concept of auto-selection systems that allows for easy and robust propagation of host cells deleted in essential genes in supplemented media before being transformed with rescuing plasmids. With that, our approach is favorable over commonly used selection strategies and has major advantage over other auto-selection systems. Our approach complements the auto-selection toolbox already available for S. cerevisiae, thus contributing a novel system that enables the use of complex peptone-based media for protein expression and metabolic engineering approaches. We therefore expect that this new strategy will be of general interest to the yeast research community in academia and industry.

Item Type: Article
Erschienen: 2021
Creators: Höler, Sebastian ; Bertl, Adam ; Degreif, Daniel
Title: Novel auto-selection systems for transformation selection of Saccharomyces cerevisiae in rich complex media
Language: English
Abstract:

The most widely used strategy for selection of yeast transformed with episomal plasmids comprises the use of auxotrophic yeast strains in combination with vectors containing complementing prototrophic marker genes. Another approach uses heterologous genes or cassettes which, if present in the vector, render the otherwise sensitive yeast strain resistant to antibiotics. In addition, auto-selection systems for Saccharomyces cerevisiae have been developed that eliminate the requirement for synthetic drop-out media or the use of antibiotics for transformation selection and subsequent plasmid maintenance in expression cultures. Here we describe a combination of host strain and vector system introducing a novel concept of auto-selection systems that allows for easy and robust propagation of host cells deleted in essential genes in supplemented media before being transformed with rescuing plasmids. With that, our approach is favorable over commonly used selection strategies and has major advantage over other auto-selection systems. Our approach complements the auto-selection toolbox already available for S. cerevisiae, thus contributing a novel system that enables the use of complex peptone-based media for protein expression and metabolic engineering approaches. We therefore expect that this new strategy will be of general interest to the yeast research community in academia and industry.

Journal or Publication Title: FEMS yeast research
Volume of the journal: 21
Issue Number: 5
Divisions: 10 Department of Biology
10 Department of Biology > Yeast Membrane Biology
Date Deposited: 12 Jul 2021 12:18
DOI: 10.1093/femsyr/foab039
Identification Number: pmid:34232310
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